Development and evaluation of novel quantitative PCR (qPCR) and loop-mediated isothermal amplification (LAMP) assays for bovine adenovirus type 7
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Herein, we report the development of molecular assays for the specific detection of bovine adenovirus type 7 (BAV-7), a prevalent pathogen associated with bovine respiratory disease (BRD). To overcome the limitations of the current diagnostic methods, we developed and optimized a TaqMan quantitative polymerase chain reaction (qPCR) assay with a limit of detection (LOD) of 50 copies per reaction (10 copies/µL) and 100% analytical specificity, showing no cross-reactivity with nine other viral pathogens commonly linked to BRD. Additionally, we designed a fluorescent quantitative loop-mediated isothermal amplification (qLAMP) assay demonstrating equivalent LOD and specificity. Both assays were evaluated using 24 bovine nasal swabs from BRD animals, revealing a 92% agreement between qPCR and qLAMP results. These assays provide powerful tools for advancing our understanding of BAV-7 epidemiology and improving disease management strategies. The qLAMP assay, with its rapid and isothermal amplification, presents potential for future development as a field-deployable diagnostic tool to enable timely detection of BAV-7 in cattle.
